Bottom Line:
Through real time polymerase chain reaction (PCR) analyses we determined that the HSFY family is largely expanded in cattle (∼70 copies) compared with human (2 functional copies, 4 HSFY-similar copies).Unexpectedly, we found that it does not vary among individual bulls as a copy number variant (CNV).HSFY expression in cattle appears restricted to the testis and its mRNA correlates positively with mRNA markers of spermatogonial and spermatocyte cells (UCHL1 and TRPC2, respectively) which suggests that HSFY is expressed (at least in part) in early germ cells.

ABSTRACTHeat shock transcription factor, Y-linked (HSFY) is a member of the heat shock transcriptional factor (HSF) family that is found in multiple copies on the Y chromosome and conserved in a number of species. Its function still remains unknown but in humans it is thought to play a role in spermatogenesis. Through real time polymerase chain reaction (PCR) analyses we determined that the HSFY family is largely expanded in cattle (∼70 copies) compared with human (2 functional copies, 4 HSFY-similar copies). Unexpectedly, we found that it does not vary among individual bulls as a copy number variant (CNV). Using fluorescence in situ hybridization (FISH) we found that the copies are dispersed along the long arm of the Y chromosome (Yq). HSFY expression in cattle appears restricted to the testis and its mRNA correlates positively with mRNA markers of spermatogonial and spermatocyte cells (UCHL1 and TRPC2, respectively) which suggests that HSFY is expressed (at least in part) in early germ cells.

pone-0017790-g004: Gel electrophoresis of the real time PCR product of both male and female DNA samples for HSFY and ZAR1.HSFY shows male-specific amplification indicating that the target genes are on the Y chromosome. Zygote arrest 1 (ZAR1) is the autosomal control gene which shows amplification in both the male and female sample. N) Negative control M) 100 bp ladder – the top and bottom band represent 200 and 100 bp, respectively (Invitrogen Canada Inc.).

Mentions:
Primer efficiencies were tested and were not found to be significantly different among targets (HSFY8, HSFY10, HSFY16) and the reference gene (SRY). We used the HSFY primers on both male and female DNA samples and as expected we found amplification in the male samples only whereas ZAR1, which is an autosomal gene, was present in both (Figure 4).

pone-0017790-g004: Gel electrophoresis of the real time PCR product of both male and female DNA samples for HSFY and ZAR1.HSFY shows male-specific amplification indicating that the target genes are on the Y chromosome. Zygote arrest 1 (ZAR1) is the autosomal control gene which shows amplification in both the male and female sample. N) Negative control M) 100 bp ladder – the top and bottom band represent 200 and 100 bp, respectively (Invitrogen Canada Inc.).

Mentions:
Primer efficiencies were tested and were not found to be significantly different among targets (HSFY8, HSFY10, HSFY16) and the reference gene (SRY). We used the HSFY primers on both male and female DNA samples and as expected we found amplification in the male samples only whereas ZAR1, which is an autosomal gene, was present in both (Figure 4).

Bottom Line:
Through real time polymerase chain reaction (PCR) analyses we determined that the HSFY family is largely expanded in cattle (∼70 copies) compared with human (2 functional copies, 4 HSFY-similar copies).Unexpectedly, we found that it does not vary among individual bulls as a copy number variant (CNV).HSFY expression in cattle appears restricted to the testis and its mRNA correlates positively with mRNA markers of spermatogonial and spermatocyte cells (UCHL1 and TRPC2, respectively) which suggests that HSFY is expressed (at least in part) in early germ cells.

ABSTRACTHeat shock transcription factor, Y-linked (HSFY) is a member of the heat shock transcriptional factor (HSF) family that is found in multiple copies on the Y chromosome and conserved in a number of species. Its function still remains unknown but in humans it is thought to play a role in spermatogenesis. Through real time polymerase chain reaction (PCR) analyses we determined that the HSFY family is largely expanded in cattle (∼70 copies) compared with human (2 functional copies, 4 HSFY-similar copies). Unexpectedly, we found that it does not vary among individual bulls as a copy number variant (CNV). Using fluorescence in situ hybridization (FISH) we found that the copies are dispersed along the long arm of the Y chromosome (Yq). HSFY expression in cattle appears restricted to the testis and its mRNA correlates positively with mRNA markers of spermatogonial and spermatocyte cells (UCHL1 and TRPC2, respectively) which suggests that HSFY is expressed (at least in part) in early germ cells.